A stable dual emulsion of Pleurotus ostreatus protein particles, its preparation method and application

Abstract

This invention discloses a stable double emulsion of oyster mushroom protein particles, its preparation method, and its applications. The preparation method first uses a pH cycling method to prepare oyster mushroom protein particles. Then, the suspension of oyster mushroom protein particles is mixed with edible oil, controlling the mass ratio of the aqueous phase to the oil phase at 20:80–90:10. After a further homogenization step, a double emulsion stable with oyster mushroom protein particles is obtained. The process conditions of this invention are mild, naturally safe, and allow for rapid continuous production, possessing industrial and large-scale application value in the fields of food, pharmaceutical, and cosmetic processing technologies.

Description

Technical Field

[0001] This invention relates to the field of food processing, and in particular to a stable double emulsion of oyster mushroom protein particles, its preparation method, and its application. Background Technology

[0002] Modern food industry and consumer demands place increasingly higher demands on the natural, healthy, and sustainable attributes of food and its ingredients. Therefore, food researchers have recently focused on exploring natural emulsifiers to replace synthetic and semi-synthetic emulsifiers, emphasizing the development of a natural, green, nutritious, healthy, and sustainable modern food industry. Protein particles, with hydrophobic and hydrophilic amino acid side chains, are typical amphiphilic particles that can spontaneously migrate to the oil-water interface, thereby reducing the interfacial tension of the emulsion during preparation and forming a mechanically strong protective layer around the droplets. With changing dietary habits, animal-derived proteins are being partially replaced by plant and edible fungal proteins. Oyster mushroom protein is a nutritious edible fungal protein with excellent food processing performance. After pH cycling treatment, amphiphilic protein particles can be obtained, which can be widely used as a safe, healthy, and sustainable emulsion stabilizer in the food industry.

[0003] Oyster mushroom protein is an edible fungus protein source with extremely high nutritional value and biological functional properties. The essential amino acid content of oyster mushroom protein meets the WHO recommended reference values, making it an excellent source of dietary protein. Oyster mushroom protein is extremely rich in glutamic acid and aspartic acid residues, which can play important roles in immune and hormonal regulation. The amphiphilic nature of oyster mushroom protein makes its emulsification application in the food industry promising. Currently, there are few reports, both domestically and internationally, on the application of oyster mushroom protein particles as emulsifiers in the food industry.

[0004] W1 / O / W2 dual emulsion refers to O / W2 droplets containing W1 / O droplets. This unique multi-chamber structure allows it to partially replace oils in the preparation of low-fat foods, and can also simultaneously encapsulate, preserve, transport, and control the release of water-soluble and fat-soluble substances.

[0005] However, dual emulsions are currently difficult to apply on a large scale in the food industry. The main reasons include the difficulty in efficiently preparing stable dual emulsions and the fact that the emulsifiers used in their preparation are mostly small-molecule surfactants or chemically modified block copolymers, while safe and environmentally friendly food-grade emulsifiers are relatively scarce. Dual emulsions can be prepared by one-step homogenization, two-step homogenization, or microfluidic devices. Microfluidic devices can produce dual emulsions with uniform particle size and high internal aqueous phase content, but the production efficiency is low and cannot meet the needs of large-scale applications in the food industry. Two-step homogenization can cause shearing and breakage of the internal droplets during the secondary homogenization process, leading to the leakage of the encapsulated material into the continuous phase, which is detrimental to the stability of the encapsulation and delivery system. One-step homogenization can greatly ensure the structural stability of the dual emulsion while maintaining high production efficiency, making it more valuable for large-scale production in the food industry and a research hotspot in the field of dual emulsions. Furthermore, dual emulsions share common storage instability behaviors such as internal aqueous phase loss; storage stability is also a key factor that must be considered in the development of dual emulsion preparation methods.

[0006] Regarding the selection of emulsifiers, to obtain stable dual emulsions, high-concentration PGPR (polyglycerol ricinoleate), Span 80 (sorbitan oleate), and other small-molecule surfactants or chemically modified inorganic particles are currently commonly used as emulsifiers. However, their potential toxic side effects do not meet current consumer demands for health and food safety. Among these, particles possess excellent irreversible adsorption properties, offering significant advantages in emulsion storage stability compared to other emulsifiers, and the development of food-grade granular emulsifiers is feasible. In summary, constructing natural, green, and sustainable food-grade granular emulsifiers that can prepare and stabilize dual emulsions via a one-step homogenization method is of great significance for the large-scale application of dual emulsions in the food industry, but it also presents a significant challenge. Typically, using only food-grade protein particles as emulsifiers is insufficient for preparing dual emulsions, resulting in emulsions with primarily traditional single-chamber emulsion structures. For example, the article "Structural and functional properties of perilla protein isolate extracted from oilseed residues and its utilization in Pickering emulsions" (doi:10.1016 / j.foodhyd.2020.106412) compared perilla protein isolates extracted by isoelectric point precipitation after three different defatting processes. The hydrophobicity values, from lowest to highest, were 101.03, 112.68, and 119.29. The nanoparticles obtained after pH cycling and homogenization of the protein were used as emulsifiers to prepare a Pickering single-chamber emulsion, rather than a double emulsion.

[0007] Oyster mushrooms and other edible fungi are delicious and highly nutritious, making them promising protein sources in the food industry. Studies have found that hydrolysates of oyster mushroom protein exhibit antioxidant activity by reducing the generation of reactive oxygen species and increasing the activity of antioxidant enzymes, demonstrating strong neuroprotective effects. Nutritionally, the essential amino acid content and amino acid score of oyster mushroom protein meet the reference values ​​recommended by FAO / WHO, with Asp and Glu being the most abundant, potentially playing a positive role in immune stimulation and endocrine regulation. In terms of physicochemical properties, the protein has an isoelectric point of approximately 4.8, a surface hydrophobicity of 113.1, and an EAI of approximately 200m. 2 / g. Summary of the Invention

[0008] The purpose of this invention is to overcome the shortcomings and deficiencies of existing technologies and provide a method for preparing a stable double emulsion of oyster mushroom protein particles. This invention obtains a stable double emulsion of protein particles from a mixture of oyster mushroom protein particles and sunflower seed oil via a one-step homogenization method, exhibiting good stability.

[0009] Another object of the present invention is to provide a double emulsion prepared by the above preparation method.

[0010] Another object of the present invention is to provide the application of the above-mentioned dual emulsion.

[0011] The objective of this invention is achieved through the following technical solution:

[0012] A method for preparing a stable dual emulsion of Pleurotus ostreatus protein particles includes the following steps:

[0013] (1) Add the oyster mushroom protein to water, adjust the pH, and stir to obtain a protein solution;

[0014] (2) Adjust the pH of the protein solution in step (1) and continue stirring;

[0015] (3) Add the oil to the solution in step (2) and homogenize to obtain a stable double emulsion of mushroom protein particles.

[0016] The mass fraction of *Pleurotus ostreatus* protein powder in the protein solution described in step (1) is 1-5%; preferably 2%.

[0017] The oyster mushroom protein mentioned in step (1) is a protein extracted from the fruiting body of oyster mushroom.

[0018] The pH adjustment mentioned in step (1) is to adjust to 10-13; preferably to 12.

[0019] The stirring conditions described in step (1) are 1000-2000 r / min for 10-20 min; preferably 1000 r / min for 15 min.

[0020] The pH adjustment in step (2) is to adjust to 2-9; preferably to 3-7; more preferably to 7.

[0021] The stirring conditions described in step (2) are 1000-2000 r / min for 10-20 min; preferably 1000 r / min for 10 min.

[0022] The oil mentioned in step (3) is at least one of animal oil and vegetable oil; preferably sunflower seed oil.

[0023] The mass ratio of oil to solution in step (3) is 1:9 to 8:2; preferably 1:1 to 7:3.

[0024] The homogenization conditions described in step (3) are: 5000-30000 r / min for 3-5 min; preferably 10000-20000 r / min for 3-4 min; more preferably 20000 r / min for 3 min.

[0025] The above preparation method yields a stable double emulsion of mushroom protein particles.

[0026] Applications of the aforementioned stable Oyster Mushroom protein particle dual emulsion in dairy products, mayonnaise, flavoring substances, bioactive substances, and drug delivery emulsions.

[0027] Specifically, the applications are as follows: when industrially producing dairy products, the technical methods of this invention can be used to produce dairy products that are free of synthetic or semi-synthetic surfactants and rich in protein; when industrially producing flavor substances, bioactive substances and drug delivery emulsions, the technical methods of this invention can be used to produce emulsions that are free of synthetic or semi-synthetic surfactants and simultaneously loaded with water-soluble and oil-soluble active substances.

[0028] The present invention has the following advantages over the prior art:

[0029] This invention ingeniously utilizes the amphiphilicity and emulsifying ability of natural emulsifier, *Pleurotus ostreatus* protein particles, to prepare a dual emulsion through a simple one-step homogenization process. The *Pleurotus ostreatus* protein particles in this invention possess both hydrophobic and hydrophilic amino acid side chains, making them typical amphiphilic particles capable of spontaneously migrating to the oil-water interface. This reduces the interfacial tension of the emulsion during preparation and forms a mechanically strong protective layer around the droplets. Utilizing this property, the emulsion is prepared by homogenizing a suspension of *Pleurotus ostreatus* protein particles with edible oil.

[0030] The present invention has the following advantages over the prior art:

[0031] 1. The overall preparation process of this invention is simple, requiring only one homogenization step to prepare the dual emulsion, avoiding the rupture of the outer droplets and leakage of the inner droplets caused by secondary homogenization shearing.

[0032] 2. The raw materials of this invention are all natural, safe and healthy, and no other synthetic emulsifiers need to be added; the product is a double emulsion with good stability.

[0033] 3. This invention cleverly utilizes the amphiphilicity and emulsifying ability of oyster mushroom protein particles to provide a novel all-natural emulsifier that can be used to prepare dual emulsions.

[0034] 4. The process conditions of this invention are simple and mild, and do not involve toxic or harmful reagents or synthetic or semi-synthetic surfactants. It is natural, green, safe and healthy. It can be used for rapid and continuous production and can be applied in the food, cosmetics and pharmaceutical industries. It has industrial and large-scale application value. Attached Figure Description

[0035] Figure 1 The image shows the appearance of emulsions with different oil contents in Example 1.

[0036] Figure 2 This is a microstructure diagram of emulsions with different oil contents in Example 1.

[0037] Figure 3 This is a particle size distribution diagram of emulsions with different oil contents in Example 1.

[0038] Figure 4 The apparent viscosity diagram shows the emulsions with different oil contents in Example 1.

[0039] Figure 5 This is an appearance diagram of different protein particle pH emulsions from Example 2.

[0040] Figure 6 This is a microstructure diagram of the pH emulsions of different protein particles in Example 2.

[0041] Figure 7 This is a particle size distribution diagram of different protein particle pH emulsions in Example 2.

[0042] Figure 8 The image shows the appearance of the emulsion after long-term storage at different oil contents in Example 3.

[0043] Figure 9 This is a microstructure diagram of the emulsions from Example 3 after long-term storage with different oil contents.

[0044] Figure 10 This is a particle size distribution diagram of the emulsion after long-term storage at different oil contents in Example 3.

[0045] Figure 11 The image shows the appearance of the emulsion after long-term storage at different pH values ​​in Example 4.

[0046] Figure 12 This is a microstructure diagram of the emulsion after long-term storage at different pH values ​​in Example 4.

[0047] Figure 13 This is a particle size distribution diagram of the emulsion after long-term storage at different pH levels in Example 4. Detailed Implementation

[0048] The present invention will be further described in detail below with reference to the embodiments and accompanying drawings, but the embodiments of the present invention are not limited thereto.

[0049] Unless otherwise specified in the following implementation plan, the test conditions are generally as per standard test conditions or the test conditions recommended by the reagent company. Unless otherwise specified, all materials and reagents used are commercially available.

[0050] Example 1

[0051] (1) Fresh oyster mushrooms were purchased from the local market of South China Agricultural University. The fruiting bodies were dried at 60℃ for 6 hours (using a food dehydrator FGJ-16, Demask, Germany) until brittle, then ground into powder (using a high-speed pulverizer ML-800Y, Moling, China) and sieved through a 70-mesh sieve. The oyster mushroom powder was defatted with n-hexane (1:4 w / v; Tianjin Fuyu Chemical, China) at room temperature. The defatted powder was dispersed in water (1:10 w / v), and the pH was adjusted to 8.0 with 2M NaOH solution. After stirring at room temperature for 2 hours, the mixture was centrifuged at 10000 r / min and 25℃ for 20 min (Eppendorf 5804R, Germany), and the supernatant was collected. The pH of the supernatant was adjusted to 4.5 with 2M HCl solution, and the mixture was centrifuged at 6000 r / min and 25℃ for 15 min, and the precipitate was collected. After redissolving the precipitate in water (1:7 w / v), the pH was adjusted to 7.0 with 2M NaOH solution. Then, the precipitate was dialyzed at 4°C for 48 h using a 7 kDa dialysis bag. After dialysis, the oyster mushroom protein powder was obtained by freeze-drying (Lyophilizer FD-10–50, Biocool, China) for 48 h.

[0052] (2) Weigh a certain amount of mushroom protein powder and disperse it in deionized water. Adjust the pH to 12.0 with 2M NaOH solution. Stir at 1000r / min at room temperature until the protein powder dissolves and continue for 15min.

[0053] (3) The pH of the above solution was adjusted back to 7.0 with 2M HCl solution, and stirred at 1000r / min for 10min at room temperature to obtain a protein particle suspension of 2.0wt%.

[0054] (4) Mix the oyster mushroom protein particle suspension and sunflower seed oil at mass ratios of 90:10 (10%), 80:20 (20%), 70:30 (30%), 60:40 (40%), 50:50 (50%), 40:60 (60%), 30:70 (70%), and 20:80 (80%), respectively, and then process them with a high-speed homogenizer at 20,000 r / min for 3 min to obtain a stable oyster mushroom protein particle double emulsion with an oil content of 10-80%.

[0055] The prepared dual emulsion was then characterized. The viscosity of the emulsion was measured using an Anton Paar MCR502 modular intelligent advanced rheometer with the following parameters: plate size 50 mm, interval 1 mm, temperature 25 °C, and shear rate 1–50 s⁻¹. -1 The microstructure of the emulsion was observed using a 40x optical microscope. The particle size distribution of the emulsion was determined using a laser particle size analyzer (Mastersizer 3000, Malvern). The refractive index of sunflower seed oil was 1.47, the dispersant was water with a refractive index of 1.33, and the test temperature was 25℃. The average particle size (μm) and particle size distribution map were obtained. The microstructure and particle size distribution of the emulsion at room temperature were analyzed, and the results are as follows: Figures 1-4 As shown.

[0056] Figure 1 The image shows the appearance of emulsions containing different oil contents in Example 1. Figure 1 It can be seen that emulsions with an oil content of 10-70% are milky white and do not exhibit oil separation, while oil-water mixtures with an oil content of 80% do not form emulsions.

[0057] The microscopic images of the emulsion in this embodiment were obtained using an optical microscope. Figure 2 These are microscopic images of emulsions containing different oil contents from Example 1. Figure 2 It can be seen that double emulsions can be prepared under conditions of oil content of 10-70%, and the internal aqueous phase increases with the increase of oil content. Double emulsions with oil content of 50-70% have a higher internal aqueous phase content.

[0058] In this embodiment, the emulsion particle size was measured using a laser particle size analyzer. Figure 3 This is a particle size distribution diagram of emulsions containing different oil contents in Example 1. Figure 3It can be seen that emulsions with uniform particle size distribution can be prepared under conditions of oil content of 10% to 70%. The particle sizes corresponding to double emulsions with oil contents of 10%, 20%, 30%, 40%, 50%, 60%, and 70% are 13.20 μm, 16.48 μm, 20.83 μm, 23.50 μm, 25.54 μm, 23.21 μm, and 28.44 μm, respectively.

[0059] Figure 4 The above diagram shows the apparent viscosity of emulsions with different oil contents in Example 1. Figure 4 It can be seen that emulsions with an oil content of 50-70% exhibit shear-thinning characteristics, while emulsions with an oil content of 10-40% show shear-thinning characteristics at low shear rates of 10 s. -1 At this point, the apparent viscosity showed a slight decrease, and the emulsion exhibited shear-thinning characteristics, which may be related to the weak interactions between emulsion droplets. Further increasing the shear rate to 10–30 s⁻¹... -1 An increase in apparent viscosity was observed, and the emulsion exhibited shear thickening behavior. When the shear rate increased to 30–50 s⁻¹, the viscosity increased further. -1 The apparent viscosity remains almost unchanged.

[0060] As can be seen from the preparation method in Example 1, the raw materials and reagents used in this invention are natural, green and safe, and the processing is simple and easy to carry out rapid and continuous production.

[0061] Example 2

[0062] (1) Weigh a certain amount of the mushroom protein powder prepared in Example 1 and disperse it in deionized water. Adjust the pH to 12.0 and stir at 1000 r / min for 15 min at room temperature until there is no precipitate at the bottom and it is completely dissolved.

[0063] (2) The pH of the above solution was adjusted to 3.0, 5.0 and 7.0 respectively using 2M HCl solution, and stirred at 1000r / min for 10min at room temperature to obtain a protein particle suspension of 2%wt.

[0064] (3) Mix the suspension of mushroom protein particles with edible oil at a water phase to oil phase mass ratio of 50:50, and then process it with a homogenizer at 20000r / min for 3min to obtain a double emulsion stabilized by mushroom protein particles.

[0065] The prepared dual emulsions were then characterized using the method described in Example 1. The microstructure of each emulsion was observed under a microscope; the particle size distribution of each emulsion was measured using a particle size analyzer. The microstructure and particle size distribution of each emulsion at room temperature were analyzed, and the results are as follows: Figures 5-7 As shown.

[0066] Figure 5The image shows the appearance of emulsions with different protein particle pH values ​​in Example 2. Figure 5 It can be seen that under the condition of 50% oil content, the emulsions with pH of 3.0, 5.0 and 7.0 are milky white, the emulsions are fine and uniform, and no emulsion separation occurs.

[0067] The microscopic images of the emulsion in this embodiment were obtained using an optical microscope. Figure 6 These are microscopic images of emulsions with different protein particle pH values ​​from Example 2. Figure 6 It can be seen that, under the condition of 50% oil content, the emulsions with pH of 3.0, 5.0 and 7.0 have uniform particle size and all emulsions contain an internal aqueous phase.

[0068] In this embodiment, the emulsion particle size was measured using a laser particle size analyzer. Figure 7 This is a particle size distribution diagram of different protein particle pH emulsions in Example 2, from... Figure 7 It can be seen that, under the condition of 50% oil content, the emulsions with pH values ​​of 3.0, 5.0 and 7.0 have uniform particle size distribution, and the particle sizes of the emulsions with pH values ​​of 3.0, 5.0 and 7.0 are 24.46 μm, 26.36 μm and 21.61 μm, respectively.

[0069] As can be seen from the preparation method in Example 2, the processing method of the present invention is simple and convenient. Multiple pH emulsions can be prepared through simple processes, which can meet the industrial and large-scale production of foods with multiple pH requirements.

[0070] Example 3

[0071] The stable *Pleurotus ostreatus* protein particle dual emulsions with an oil content of 10-80% prepared in Example 1 were stored at room temperature for 30 days. The microstructure of each emulsion was observed under a microscope at 1, 5, 10, and 30 days. The particle size distribution of each emulsion was measured using a particle size analyzer at 1, 3, 5, 10, 15, 20, and 30 days. The changes in microstructure and particle size distribution of each emulsion after storage at room temperature were analyzed. The results are as follows: Figures 8-10 As shown.

[0072] Figure 8 The images shown are of the emulsion appearance after long-term storage at different oil contents in Example 3. Figure 8 It can be seen that fresh emulsions with an oil content of 10-70% do not exhibit oil separation, emulsions with an oil content of 10-20% show oil separation after 10 days of storage at room temperature, emulsions with an oil content of 10-20% show obvious oil separation after 30 days of storage, and emulsions with an oil content of 30-70% show slight oil separation.

[0073] The microscopic images of the emulsion in this embodiment were obtained using an optical microscope. Figure 9 The image shows the microstructure of the emulsions from Example 3 after long-term storage at different oil contents. Figure 9 It was found that all emulsions contained an internal aqueous phase. Emulsions with an oil content of 20-70% showed relatively small changes in particle size distribution during 30 days of storage. Among them, emulsions with an oil content of 30-50% showed the smallest change in particle size after 30 days of storage, followed by emulsions with an oil content of 60%. Emulsions with an oil content of 70% showed a similar change in particle size after 20 days of storage as emulsions with an oil content of 30-50%, with almost no change in particle size. Only a small change in particle size was observed after 30 days of storage. This indicates that the dual Pickering emulsions stabilized by *Pleurotus ostreatus* protein particles prepared by the one-step method have excellent storage stability. The particle size of emulsions with an oil content of 20-60% increased with increasing oil content after 30 days of storage, showing a similar trend to the particle size of freshly prepared emulsions. This indicates that the present invention can obtain dual emulsions with the desired particle size by adjusting the oil content.

[0074] In this embodiment, the emulsion particle size was measured using a laser particle size analyzer. Figure 10 This is a particle size distribution diagram of the emulsion after long-term storage at different oil contents in Example 3. Figure 10 It was found that the particle size distribution of emulsions with oil content of 20-70% showed little change during 30 days of storage. Specifically, the particle size ranges of emulsions with oil content of 30% and 50% after 30 days of storage were 21.01-21.63 μm and 25.55-26.20 μm, respectively, with a particle size change of less than 1 μm. The particle size range of the emulsion with oil content of 40% during storage of 3-30 days was 25.15-25.57 μm, with a particle size change of less than 1 μm. The particle size range of the emulsion with oil content of 60% after 30 days of storage was 23.68-25.17 μm, with a particle size change of approximately 1.5 μm. This indicates that the double Pickering emulsion stabilized by *Pleurotus ostreatus* protein particles prepared by the one-step method has excellent storage stability. After 30 days of storage, the multi-compartment structure was still clearly observed, and the number of internal droplets did not decrease significantly.

[0075] As can be seen from the storage stability analysis of Example 3, the emulsions prepared with various oil contents involved in this invention have excellent storage stability and provide a wide range of adjustable oil contents, which has great application prospects in the industrial and large-scale production of food, cosmetics and pharmaceuticals.

[0076] Example 4

[0077] The stable *Pleurotus ostreatus* protein particle dual emulsions with pH 3–7 prepared in Example 2 were stored at room temperature for 30 days. The microstructure of each emulsion was observed under a microscope at 1, 5, 10, and 30 days. The particle size distribution of each emulsion was measured using a particle size analyzer at 1, 5, 10, 15, 20, and 30 days. The changes in microstructure and particle size distribution of each emulsion after storage at room temperature were analyzed. The results are as follows: Figures 11-13 As shown.

[0078] Figure 11 These are the appearance images of the emulsions after long-term storage at different pH levels in Example 4. Figure 11 It was found that all emulsions exhibited slight emulsification, with no oil separation observed. Emulsions with pH values ​​of 7.0 and 5.0 showed the least degree of emulsification, indicating their suitability for neutral and weakly acidic food production. The degree of emulsification did not increase after 1–30 days of storage, demonstrating excellent storage stability.

[0079] The microscopic images of the emulsion in this embodiment were obtained using an optical microscope. Figure 12 These are microstructure diagrams of emulsions stored for a long period at different pH levels in Example 4. Figure 12 It can be seen that all emulsions contain an internal aqueous phase. During the storage period of 1 to 30 days, the particle size distribution of all emulsions is uniform. Among them, the particle size of the emulsion with pH 7.0 is the smallest, and the particle size of the emulsion with pH 5.0 is the largest.

[0080] In this embodiment, the emulsion particle size was measured using a laser particle size analyzer. Figure 13 This is a particle size distribution diagram of the emulsion after long-term storage at different pH levels in Example 4. Figure 13 It was found that all emulsions maintained a uniform particle size distribution after 30 days of storage. The particle size range of the emulsion at pH 3.0 was 24.46–25.18 μm, with a variation of less than 1 μm; the particle size range of the emulsion at pH 5.0 was 26.36–29.66 μm, with a variation of approximately 3.3 μm; and the particle size range of the emulsion at pH 7.0 was 21.61–24.46 μm, with a variation of approximately 2.8 μm. This indicates that the emulsions at different pH values ​​all exhibited excellent storage stability.

[0081] As can be seen from the storage stability analysis of Example 4, the emulsions prepared at various pH values ​​involved in this invention all have excellent storage stability. In the production of emulsion-type products with multiple pH requirements, this invention provides a wide pH adjustment range and has great application prospects in the industrial and large-scale production of food, cosmetics and pharmaceuticals.

[0082] The above embodiments are preferred embodiments of the present invention, but the embodiments of the present invention are not limited to the above embodiments. Any changes, modifications, substitutions, combinations, or simplifications made without departing from the spirit and principle of the present invention shall be considered equivalent substitutions and shall be included within the protection scope of the present invention.

Claims

1. A method for preparing a stable dual emulsion of Pleurotus ostreatus protein particles, characterized in that... Includes the following steps: (1) Add the oyster mushroom protein to water, adjust the pH, and stir to obtain a protein solution; (2) Adjust the pH of the protein solution in step (1) and continue stirring; (3) Add the oil to the solution in step (2), homogenize, and obtain a stable double emulsion of mushroom protein particles; The mass fraction of *Pleurotus ostreatus* protein powder in the protein solution described in step (1) is 5%; The pH adjustment mentioned in step (1) is to adjust it to 10-13; The stirring conditions described in step (1) are 1000-2000 r / min for 10-20 min; The pH adjustment mentioned in step (2) is to adjust it to 2-9; The stirring conditions described in step (2) are 1000-2000 r / min for 10-20 min; The oil mentioned in step (3) is sunflower seed oil; The mass ratio of oil to solution in step (3) is 1:1 to 7:3; The homogenization conditions described in step (3) are 20,000 r / min for 3 min.

2. The stable Pleurotus ostreatus protein particle dual emulsion prepared by the preparation method according to claim 1.

3. The use of the double emulsion of mushroom protein particles as described in claim 2 in dairy products, mayonnaise, flavoring substances, bioactive substances, and drug delivery emulsions.

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